doubt about Detoxi-Gel™ Endotoxin Removing Gel

Discussion of all aspects of cellular structure, physiology and communication.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
jasmina
Garter
Garter
Posts: 37
Joined: Fri Sep 12, 2008 1:54 am
Contact:

doubt about Detoxi-Gel™ Endotoxin Removing Gel

Post by jasmina » Fri Jan 08, 2010 4:41 am

Hi there
these days i will purify my samples with Detoxi-Gel™ Endotoxin Removing Gel, but i could not understand protocol very well.
1 with a gravity-flow column, the sample will begin to emerge from the column about 90% of the bed volumn has been collected. this means after apply sample to column, there is 10% sample loss, isn't it?
2 The Pierce 1ml Detoxi-GEL column has a void volumn of 0.3-0.5ml. this means after apply sample to column, there is 0.3-0.5ml sample detained in column, right?
these 2 sentense could not consistent, i am confusing.
Anyone who can explan it for me ? thank you very much!

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Re: doubt about Detoxi-Gel™ Endotoxin Removing Gel

Post by JackBean » Fri Jan 08, 2010 8:33 am

jasmina wrote:these 2 sentense could not consistent, i am confusing.

these 2 sentences could not consider/understand(?), I'm confused :wink:

Well, I found only this
http://www.piercenet.com/files/0415dh5.pdf
You probably do not know, what is bed or void volume.
The void volume is volume of liquid in the column, that is, it is the volume, when comes out the non-released fraction.
So, you do not have to care about the first 0.3-0.5 ml of eluent, because there is only the buffer.
The bed volume is the total volume of the column, including the matrice, so you can actually calculate it just as the inner volume of your column.
So, basically, calculate your bed volume
http://www.novasep.com/misc/glossary.as ... =&search=B
and after 90% of this volume will get out, you should start to collect fractions ;)
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

jasmina
Garter
Garter
Posts: 37
Joined: Fri Sep 12, 2008 1:54 am
Contact:

Post by jasmina » Sat Jan 09, 2010 2:11 am

To jackbean
thank you for your reply.
but on that protocol, i could not distinguish resin-bed volume and bed volume, what's differece? And i checked column just now, the column is graduated (maximum scale is 2ml), can i take it as column volume?
shamed to tell you that i could not understand your last sentense :oops: . why should i collect fractions after 90% of this volumn got out, rather than 0.3-0.5ml got out?
thank you in advance!

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Post by JackBean » Sat Jan 09, 2010 9:35 am

IMHO, the resin-bed volume is the volume of the resin in the column, whereas bed volume is the whole volume of the colon (that is volume of resin + volume of liquid)
What do you mean by graduated column?

Well, you had two sentences in your first post, in first you had 0.3-0.5 ml and in second you had 90% ;)
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

jasmina
Garter
Garter
Posts: 37
Joined: Fri Sep 12, 2008 1:54 am
Contact:

Post by jasmina » Mon Jan 11, 2010 5:59 am

To Jackbean
graduated column means the column was marked, so there is no need to calculate the bed volume.
I use same column, why detained buffer volume is different in these 2 sentense ? :?:

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Post by JackBean » Mon Jan 11, 2010 8:42 am

I don't know :) Isn't the second sentence about using some pump?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

jasmina
Garter
Garter
Posts: 37
Joined: Fri Sep 12, 2008 1:54 am
Contact:

Post by jasmina » Mon Jan 11, 2010 9:06 am

To Jackbean
I use gravity-flow column, and on this protocol, they did not mention about pump. :?: confusing

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Post by JackBean » Mon Jan 11, 2010 11:58 am

In that case, I think, you should collect your sample after 80-90% of the bed volume has left ;)
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

Post Reply

Who is online

Users browsing this forum: No registered users and 5 guests